LOCUS       sequence-255899-        8898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains mScarlet with CLASP. This consists of the plasma 
            membrane anchor (pm-LOVTRAP) and the cargo (Zdk-mScarlet-yeLANS). 
            CLASP modulates nuclear localization of mScarlet in response to bl.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8898)
  AUTHORS   Chen SY, Osimiri LC, Chevalier M, Bugaj LJ, Nguyen TH, Greenstein 
            RA, Ng AH, Stewart-Ornstein J, Neves LT, El-Samad H
  TITLE     Optogenetic Control Reveals Differential Promoter Interpretation of 
            Transcription Factor Nuclear Translocation Dynamics.
  JOURNAL   Cell Syst. 2020 Sep 4. pii: S2405-4712(20)30293-3. doi: 
            10.1016/j.cels.2020.08.009.
  PUBMED    32898473
REFERENCE   2  (bases 1 to 8898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Syst. 
            2020 Sep 4. pii: S2405-4712(20)30293-3. doi: 
            10.1016/j.cels.2020.08.009."
COMMENT     Sequence Label: pCLASP_mScarlet_CLASP_RGS2membrane
FEATURES             Location/Qualifiers
     source          1..8898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             218..1021
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(1663..2478)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1663..2478)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2386..2405
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2470..2489
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2703..3291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3192..3211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      6165..6352
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6220..6244)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 gttacagaga tgttacgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt
       61 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
      121 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
      181 gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
      241 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
      301 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
      361 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
      421 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
      481 cttcgaagac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
      541 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
      601 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
      661 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
      721 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
      781 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
      841 caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc
      901 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
      961 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     1021 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     1081 attatatcag ttattacccg agtagagcac ttgaatccac tgccccggga atctcggtcg
     1141 taatgatttc tataatgacg aaaaaaaaaa aattggaaag aaaaagcttc atggccttta
     1201 taaaaaggaa ctatccaata cctcgccaga accaagtaac agtattttac ggggcacaaa
     1261 tcaagaacaa taagacagga ctgtaaagat ggacgcattg aactccaaag aacaacaaga
     1321 gttccaaaaa gtagtggaac aaaagcaaat gaaggatttc atgcgtttgt actctaatct
     1381 ggtagaaaga tgtttcacag actgtgtcaa tgacttcaca acatcaaagc taaccaataa
     1441 ggaacaaaca tgcatcatga agtgctcaga aaagttcttg aagcatagcg aacgtgtagg
     1501 gcagcgtttc caagaacaaa acgctgcctt gggacaaggc ttgggccgat aaggtgtact
     1561 ggcgtatata tatctaatta tgtatctctg gtgtagccca tttttagcat gtaaatataa
     1621 agaccgagcg gccgcgtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag
     1681 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     1741 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     1801 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     1861 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     1921 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     1981 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgaggcgaaa
     2041 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     2101 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     2161 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     2221 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     2281 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     2341 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     2401 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc aagacgtttc
     2461 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     2521 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     2581 gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagtcat gaccaaaatc
     2641 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2701 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2761 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2821 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2881 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2941 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3001 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3061 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3121 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3181 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3241 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3301 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3361 gcgttatccc ctgattctgt ggataaccgt gcggccgcca atgctaaatt cgagtgaaac
     3421 acaggaagat cagaaaatcc tcatttcatc catattaaca ataatttcaa atgtttattt
     3481 gcattatttg aaactaggca agacaagcaa cgaaacgttt ttgaaaattt tgagtatttt
     3541 caataaattt gtagaggact cagatattga aaaaaagcta cagcaattaa tacttgataa
     3601 gaagagtatt gagaagggca acggttcatc atctcatgga tctgcacatg aacaaacacc
     3661 agagtcaaac gacgttgaaa ttgaggctac tgcgccaatt gatgacaata cagacgatga
     3721 taacaaaccg aagttatctg atgtagaaaa ggattaaaga tgctaagaga tagtgatgat
     3781 atttcataaa taatgtaatt ctatatatgt taattacctt ttttgcgagg catatttatg
     3841 gtgaaggata agttttgacc atcaaagaag gttaatgtgg ctgtggtttc agggtccata
     3901 aagcccacat ggataacatt acccctgaat tcgcatctag actgaactgg ccgataattg
     3961 cagacgaacg aagaggatgt ccaatatttt ttttaaggaa taaggatact tcaagactag
     4021 attcccccct gcattcccat cagaaccgta aaccttggcg ctttccttgg gaagtattca
     4081 agaagtgcct tgtccggttt ctgtggctca caaaccagcg cgcccgatat ggctttcttt
     4141 tcacttatga atgtaccagt acgggacaat tagaacgctc ctgtaacaat ctctttgcaa
     4201 atgtggggtt acattctaac catgtcacac tgctgacgaa attcaaagta aaaaaaaatg
     4261 ggaccacgtc ttgagaacga tagattttct ttattttaca ttgaacagtc gttgtctcag
     4321 cgcgctttat gttttcattc atacttcata ttataaaata acaaaagaag aatttcatat
     4381 tcacgcccaa gaaatcaggc tgctttccaa atgcaattga cacttcatta gccatcacac
     4441 aaaactcttt cttgctggag cttcttttaa aaaagacctc agtacaccaa acacgttacc
     4501 cgacctcgtt attttacgac aactatgata aaattctgaa gaaaaaataa aaaaattttc
     4561 atacttcttg cttttattta aaccattgaa tgatttcttt tgaacaaaac tacctgtttc
     4621 accaaaggaa atagaaagaa aaaatcaatt agaagaaaac aaaaaacaaa agatctatgg
     4681 gttctgttga taataagttt aataaagaga aaacaagagc tggagccgaa atacactctt
     4741 tgcctaactt aaatgtagaa cagaagttcg cttttatagt atctttgttt gatgatccat
     4801 cacaatctgc caatttgttg gcagaggcta agaaattaaa cgacgctcaa gcaccaaaag
     4861 gttctgtatc aaaaggcgag gcggtcatta aagagttcat gaggtttaaa gtccatatgg
     4921 aaggaagcat gaatggtcac gagtttgaga tagagggaga gggtgaggga aggccgtacg
     4981 aagggacgca aactgcgaag ttgaaggtaa cgaaaggcgg ccctctgcca ttctcttggg
     5041 acatcctttc accgcaattt atgtacggct ctagggcctt cacaaaacac cccgccgata
     5101 ttccggatta ctataaacaa agttttcctg agggcttcaa gtgggaaaga gtaatgaact
     5161 tcgaggatgg gggagctgtg acggttactc aagacactag cctggaggac ggtactctta
     5221 tatacaaggt aaaactaaga gggacgaatt ttccgccaga cggccccgtg atgcaaaaaa
     5281 aaactatggg gtgggaagca tccaccgaga ggctgtaccc cgaagacggg gtcctgaagg
     5341 gtgatattaa aatggcatta agattaaaag atggcgggag atacttagct gacttcaaga
     5401 ctacatataa agccaaaaaa cctgtacaga tgcctggagc ttacaatgtg gatcgtaagc
     5461 tggatattac atcccacaac gaagattata ccgtggtgga gcagtacgag cgttcagagg
     5521 ggcgtcactc caccgggggt atggatgaat tatacaagag cggtcttaga agcagggcgc
     5581 aagcgtcaaa ttcagcggta gatggaacag caggtccagg tagtactggc agcagaggat
     5641 cccccgaatt cggcattgat ctgagcggcc tgaccctgca gggttctgga agtgggtcgt
     5701 tggctactac acttgaacgt attgagaaga actttgtcat tactgaccca agattgccag
     5761 ataatcccat tatattcgcg tccgatagtt tcttgcagtt gacagaatat agccgtgaag
     5821 aaattttggg aagaaactgc aggtttctac aaggtcctga aactgatcgc gcgacagtga
     5881 gaaaaattag agatgccata gataaccaaa cagaggtcac tgttcagctg attaattata
     5941 caaagagtgg taaaaagttc tggaacctct ttcacttgca gcctatgcga gatcagaagg
     6001 gagatgtcca gtactttatt ggggttcagt tggatggaac tgagcatgtc cgagatgctg
     6061 ccgagagaga gggagtcatg ctgattaaga aaactgcaga aaatattgat gctgctgcta
     6121 agagatcttg gtctatggct ttcgagctcg gatcctaact cgaggcgaat ttcttatgat
     6181 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
     6241 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
     6301 tgctttctca ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcatgccga
     6361 gcaaatgcct gcaaatcgct ccccatttcg ctgccaaacc agatgtcaac acagctacaa
     6421 cgaagaggat gtccaatatt ttttttaagg aataaggata cttcaagact agattccccc
     6481 ctgcattccc atcagaaccg taaaccttgg cgctttcctt gggaagtatt caagaagtgc
     6541 cttgtccggt ttctgtggct cacaaaccag cgcgcccgat atggctttct tttcacttat
     6601 gaatgtacca gtacgggaca attagaacgc tcctgtaaca atctctttgc aaatgtgggg
     6661 ttacattcta accatgtcac actgctgacg aaattcaaag taaaaaaaaa tgggaccacg
     6721 tcttgagaac gatagatttt ctttatttta cattgaacag tcgttgtctc agcgcgcttt
     6781 atgttttcat tcatacttca tattataaaa taacaaaaga agaatttcat attcacgccc
     6841 aagaaatcag gctgctttcc aaatgcaatt gacacttcat tagccatcac acaaaactct
     6901 ttcttgctgg agcttctttt aaaaaagacc tcagtacacc aaacacgtta cccgacctcg
     6961 ttattttacg acaactatga taaaattctg aagaaaaaat aaaaaaattt tcatacttct
     7021 tgcttttatt taaaccattg aatgatttct tttgaacaaa actacctgtt tcaccaaagg
     7081 aaatagaaag aaaaaatcaa ttagaagaaa acaaaaaaca aaagatctat gaagagaaca
     7141 ttgttgaagg attggaaaac cagattatca tacttcttac agaactcatc tacacctggt
     7201 aagccaaaga caggtaagaa atctaaacaa caaggcggtg gcggaagtgg cggaggaggt
     7261 tcgggcggaa gtggttctgc tgaaggttct gttgctagac aaccagactt gttgacttgt
     7321 gacgacgaac caatccacat cccaggtgct atccaaccac acggtttgtt gttggctttg
     7381 gctgctgaca tgactatcgt tgctggttct gacaacttgc cagaattgac tggtttggct
     7441 atcggtgctt tgatcggtag atctgctgct gacgttttcg actctgaaac tcacaacaga
     7501 ttgactatcg ctttggctga accaggtgct gctgttggtg ctccaatcac tgttggtttc
     7561 actatgagaa aggacgctgg tttcatcggt tcttggcaca gacacgacca attgatcttc
     7621 ttggaattgg aaccaccaca aagagatgtt gctgaaccac aagctttctt cagaagaact
     7681 aactctgcta tcagaagatt gcaagctgct gaaactttgg aatctgcttg tgctgctgct
     7741 gctcaagaag ttagaaagat cactggtttc gacagagtta tgatctacag attcgcttct
     7801 gacttctctg gtgaagttat cgctgaagac agatgtgctg aagttgaatc taagttgggt
     7861 ttgcactacc cagcttctac tgttccagct caagctagaa gattgtacac tatcaaccca
     7921 gttagaatca tcccagacat caactacaga ccagttccag ttactccaga cttgaaccca
     7981 gttactggta gaccaatcga cttgtctttc gctatcttga gatctgtttc tccagttcac
     8041 ttggaattca tgagaaacat cggtatgcac ggtactatgt ctatctctat cttgagaggt
     8101 gaaagattgt ggggtttgat cgtttgtcac cacagaactc catactacgt tgacttggac
     8161 ggtagacaag cttgtgaatt ggttgctcaa gttttggctt ggcaaatcgg tgttatggaa
     8221 gaaggatcct tagctactac gctagagcgt atagaaaaaa atttcgtgat caccgatcca
     8281 agattgccgg acaacccaat catatttgct tccgattcct ttttacaact gaccgaatac
     8341 agtagagagg agattctggg taggaattgc aggttccttc agggtcccga gactgatcgt
     8401 gcaacagtta ggaaaattag agatgcgata gacaaccaga cagaagttac cgtgcagttg
     8461 atcaattata ccaagagcgg aaaaaaattc tggaacttat ttcatttaca gccaatgcgt
     8521 gaccagaagg gcgacgtaca gtatttcatt ggtgtgcaac tagacgggac agagcacgta
     8581 cgtgacgcag cagaaagaga gggcgtgatg ttgataaaaa aaactgcaga gaatatcgac
     8641 gaggctgcga aagagctata atggcattga attgaattga aatcgataga tcaatttttt
     8701 tcttttctct ttccccatcc tttacgctaa aataatagtt tattttattt tttgaatatt
     8761 ttttatttat atacgtatat atagactatt atttatcttt taatgattat taagattttt
     8821 attaaaaaaa aattcgctcc tcttttaatg cctttatgca gttttttttt cccattcgat
     8881 atttctatgt gctgagca
//
